title: Identification of novel lipolytic genes and gene families by screening of metagenomic libraries derived from soil samples of the German Biodiversity Exploratories.
abstract: Microbial metagenomes derived from soils are rich sources for the discovery of novel genes and biocatalysts. Fourteen environmental plasmid and seven fosmid libraries obtained from 10 German forest soils (A horizons) and six grassland soils (A and B horizons) were screened for genes conferring lipolytic activity. The libraries comprised approximately 29.3 Gb of cloned soil DNA. Partial activity-based screening of the constructed libraries resulted in the identification of 37 unique lipolytic clones. The amino acid sequences of the 37 corresponding lipolytic gene products shared 29-90% identity to other lipolytic enzymes, which were mainly uncharacterized or derived from uncultured microorganisms. Multiple sequence alignments and phylogenetic tree analysis revealed that 35 of the predicted proteins were new members of known families of lipolytic enzymes. The remaining two gene products represent two putatively new families. In addition, sequence analysis indicated that two genes encode true lipases, whereas the other genes encode esterases. The determination of substrate specificity and chain-length selectivity using different triacylglycerides and p-nitrophenyl esters of fatty acids as substrates supported the classification of the esterases.
authors.affiliation: Department of Genomic and Applied Microbiology, Institute of Microbiology and Genetics, Georg-August University Göttingen, Göttingen, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Nacke
authors.forename: Christiane
authors.initials: C
authors.lastname: Will
authors.forename: Sarah
authors.initials: S
authors.lastname: Herzog
authors.forename: Boris
authors.initials: B
authors.lastname: Nowka
authors.forename: Martin
authors.initials: M
authors.lastname: Engelhaupt
authors.forename: Rolf
authors.initials: R
authors.lastname: Daniel
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000145:classification / Q000201:enzymology / Q000235:genetics; D044822:Biodiversity; D004950:Esterases / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D005798:Genes, Bacterial; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008066:Lipolysis / Q000235:genetics*; D054892:Metagenome; D056186:Metagenomics; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012987:Soil / Q000737:chemistry; D012988:Soil Microbiology; D013379:Substrate Specificity
chemical_list: D012987:Soil; D004950:Esterases; D008049:Lipase